Figure 5. SLCO2B1 expression is sufficient for the uptake of heme analogs.

(A) Chemical structure of Zinc Mesoporphyrin (ZnMP).

(B-D) Flow cytometry analysis of ZnMP uptake in (B) Jurkat cells expressing vector, SLCO2A1 or SLCO2B1 cDNA, treated with 5 μM ZnMP for 24 h (C) HepG2 cells expressing vector or SLCO2B1 cDNA treated with 1 μM ZnMP for 15 min (D) PaTu-8988t cells expressing vector or SLCO2B1 cDNA treated with 2 μM ZnMP for 15 min.

(E) Dose curve for ZnMP uptake in mouse HY15549 pancreas cells expressing vector or Slco2b1 cDNA treated with indicated doses of ZnMP for 15 min at room temperature. (Mean ± SEM, n= 3, Student’s t-test, 95% confidence interval).

(F) Time course for ZnMP uptake in mouse HY15549 pancreas cells expressing vector or Slco2b1 cDNA treated with 1 μM ZnMP for the indicated time points.

(G) Chemical structures of Heme B and Hemin.

(H) Fold change in the number (log₂) of Jurkat cells expressing control vector, SLCO2A1 or SLCO2B1 cDNA, during 5-day incubation with or without Hemin treatment at indicated concentrations. (Mean ± SEM, n= 3, Student’s t-test, 95% confidence interval).

(I) Representative images of cell pellets from Hemin-treated Jurkat cells expressing vector, SLCO2A1 or SLCO2B1 cDNA. Cells were treated with 30 μM Hemin for 24 h prior to pelleting.

(J) Immunoblot analysis for ALAS1 in Succinyl Acetone (1mM) or Hemin (30 μM) treated Jurkat cells expressing vector or SLCO2B1 cDNA. β-actin was used as loading control.

See also Figure S5